Digital amplification
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Accused Products
Abstract
The identification of pre-defined mutations expected to be present in a minor fraction of a cell population is important for a variety of basic research and clinical applications. The exponential, analog nature of the polymerase chain reaction is transformed into a linear, digital signal suitable for this purpose. Single molecules can be isolated by dilution and individually amplified; each product is then separately analyzed for the presence of pre-defined mutations. The process provides a reliable and quantitative measure of the proportion of variant sequences within a DNA sample.
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Citations
108 Claims
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1-103. -103. (canceled)
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104. A molecular beacon probe comprising:
an oligonucleotide comprising a stem and a loop structure and having a photoluminescent dye at one of the 5′
or 3′
ends and a quenching agent at the opposite 5′
or 3′
end, wherein the loop consists of 14-26 bases and has a Tm of 50-51°
C., and wherein the stem consists of 4 base pairs having a sequence 5′
-CACG-3′
.- View Dependent Claims (105, 106)
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107. A molecular beacon probe comprising:
an oligonucleotide comprising a stem and a loop structure and having a photoluminescent dye at one of the 5′
or 3′
ends and a quenching agent at the opposite 5′
or 3′
end, wherein the loop consists of 14-26 bases and has a Tm of 54-56°
C., and wherein the stem consists of 4-6 base pairs comprising a sequence 5′
-CACG-3′
.
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108. A pair of molecular beacon probes comprising:
a first oligonucleotide comprising a first stem and a first loop structure and having a photoluminescent dye at one of the 5′
or 3′
ends and a quenching agent at the opposite 5′
or 3′
end, wherein the first loop consists of 14-26 bases and has a Tm of 50-51°
C., and wherein the first stem consists of 4 base pairs having a sequence 5′
-CACG-3′
; and
Specification